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  • A disposable chemical heater and dry enzyme preparation for lysis and extraction of DNA and RNA from microorganisms.

A disposable chemical heater and dry enzyme preparation for lysis and extraction of DNA and RNA from microorganisms.

Analytical methods : advancing methods and applications (2016-04-14)
J R Buser, X Zhang, S A Byrnes, P D Ladd, E K Heiniger, M D Wheeler, J D Bishop, J A Englund, B Lutz, B H Weigl, P Yager
ABSTRACT

Sample preparation, including bacterial lysis, remains a hurdle in the realization of complete point-of-care tests for many pathogens. Here, we developed a sample preparation methodology for enzymatic lysis and sample heating for low-resource, point-of-care applications. We show an instrument-free chemical heater system for rapid lysis of a gram-positive bacterium (Staphylococcus aureus) and an RNA virus (human respiratory syncytial virus) using a dried lysis enzyme mixture (achromopeptidase) for S. aureus. After a lysis step (<1 minute), lysis enzymes are heat deactivated (<5 minutes) using a simple disposable chemical heater. We demonstrated that both DNA and RNA in the heat-treated sample could be directly amplified without purification, even in the presence of a clinically-obtained human nasal sample. This simple approach to dry enzyme storage and sample heating is adaptable to many applications where samples need to be lysed, including use in low-resource laboratories and in single-use or cartridge-based point-of-care diagnostic devices.

MATERIALS
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Sigma-Aldrich
Achromopeptidase from bacteria, lyophilized powder, ≥1,000 units/mg solid